Endocrinology, Vol 136, 4271-4277, Copyright © 1995 by Endocrine Society

ARTICLES

Agonist-stimulated phosphorylation of the G protein-coupled receptor for parathyroid hormone (PTH) and PTH-related protein

E Blind, T Bambino and RA Nissenson
Endocrine Unit, Veterans Administration Medical Center, San Francisco, California 94121, USA.

The objectives of the present study were to determine whether the G protein-coupled receptor for PTH and PTH-related protein (PTHrP) is subject to agonist-specific phosphorylation and to characterize the relevant kinase(s). The opossum kidney PTH/PTHrP receptor stably expressed in human embryonic kidney 293 cells was coupled to adenylyl cyclase, with half-maximal activation occurring in the presence of 0.1 nM bovine (b) PTH-(1-34). Immunoprecipitation of extracts of 32P- labeled cells using a monoclonal antibody to the PTH/PTHrP receptor revealed the presence of a major 32P-labeled protein of approximately 85 kilodaltons that was not evident in untransfected 293 cells. bPTH-(1- 34) treatment produced a rapid dose-dependent increase in phosphorylation of the 85-kilodalton receptor, with a maximal effect that was 3.5 +/- 0.7-fold (n = 4) over basal. Half-maximal phosphorylation occurred with 10 nM bPTH-(1-34), similar to the hormone concentration required for 50% receptor occupancy. Activation of protein kinase A or protein kinase C with forskolin or phorbol 12- myristate 13-acetate also increased PTH/PTHrP receptor phosphorylation, but to a lesser degree than PTH. Neither of these kinases mediated the effect of PTH, as blockade of the protein kinase A pathway (with H-89) or the protein kinase C pathway (with the bisindolylmaleimide GF 109203X) did not inhibit bPTH-(1-34)-induced PTH/PTHrP receptor phosphorylation. These results suggest that agonist-stimulated PTH/PTHrP receptor phosphorylation may involve a nonsecond messenger- activated kinase, such as a member of the G protein-coupled receptor kinase family.

This article has been cited by other articles:

				A. Rey, D. Manen, R. Rizzoli, J. Caverzasio, and S. L. Ferrari Proline-rich Motifs in the Parathyroid Hormone (PTH)/PTH-related Protein Receptor C Terminus Mediate Scaffolding of c-Src with beta-Arrestin2 for ERK1/2 Activation J. Biol. Chem., December 15, 2006; 281(50): 38181 - 38188. [Abstract] [Full Text] [PDF]

				M. Castro, V. O. Nikolaev, D. Palm, M. J. Lohse, and J.-P. Vilardaga Turn-on switch in parathyroid hormone receptor by a two-step parathyroid hormone binding mechanism PNAS, November 1, 2005; 102(44): 16084 - 16089. [Abstract] [Full Text] [PDF]

				C. A. Syme, P. A. Friedman, and A. Bisello Parathyroid Hormone Receptor Trafficking Contributes to the Activation of Extracellular Signal-regulated Kinases but Is Not Required for Regulation of cAMP Signaling J. Biol. Chem., March 25, 2005; 280(12): 11281 - 11288. [Abstract] [Full Text] [PDF]

				W. B. Sneddon, C. E. Magyar, G. E. Willick, C. A. Syme, F. Galbiati, A. Bisello, and P. A. Friedman Ligand-Selective Dissociation of Activation and Internalization of the Parathyroid Hormone (PTH) Receptor: Conditional Efficacy of PTH Peptide Fragments Endocrinology, June 1, 2004; 145(6): 2815 - 2823. [Abstract] [Full Text] [PDF]

				S. Chauvin, M. Bencsik, T. Bambino, and R. A. Nissenson Parathyroid Hormone Receptor Recycling: Role of Receptor Dephosphorylation and {beta}-Arrestin Mol. Endocrinol., December 1, 2002; 16(12): 2720 - 2732. [Abstract] [Full Text] [PDF]

				X. Chen, J.-C. Dai, and E. M. Greenfield Termination of immediate-early gene expression after stimulation by parathyroid hormone or isoproterenol Am J Physiol Cell Physiol, November 1, 2002; 283(5): C1432 - C1440. [Abstract] [Full Text] [PDF]

				H. A. W. Tawfeek, F. Qian, and A. B. Abou-Samra Phosphorylation of the Receptor for PTH and PTHrP Is Required for Internalization and Regulates Receptor Signaling Mol. Endocrinol., January 1, 2002; 16(1): 1 - 13. [Abstract] [Full Text] [PDF]

				H. A. W. Tawfeek, J. Che, F. Qian, and A. B. Abou-Samra Parathyroid hormone receptor internalization is independent of protein kinase A and phospholipase C activation Am J Physiol Endocrinol Metab, September 1, 2001; 281(3): E545 - E557. [Abstract] [Full Text] [PDF]

				P. R. Turner, S. Mefford, S. Christakos, and R. A. Nissenson Apoptosis Mediated by Activation of the G Protein-Coupled Receptor for Parathyroid Hormone (PTH)/ PTH-Related Protein (PTHrP) Mol. Endocrinol., February 1, 2000; 14(2): 241 - 254. [Abstract] [Full Text]

				R. R. Miles, J. P. Sluka, R. F. Santerre, L. V. Hale, L. Bloem, G. Boguslawski, K. Thirunavukkarasu, J. M. Hock, and J. E. Onyia Dynamic Regulation of RGS2 in Bone: Potential New Insights into Parathyroid Hormone Signaling Mechanisms Endocrinology, January 1, 2000; 141(1): 28 - 36. [Abstract] [Full Text] [PDF]

				M. Mannstadt, H. Juppner, and T. J. Gardella Receptors for PTH and PTHrP: their biological importance and functional properties Am J Physiol Renal Physiol, November 1, 1999; 277(5): F665 - F675. [Abstract] [Full Text] [PDF]

				F. Dicker, U. Quitterer, R. Winstel, K. Honold, and M. J. Lohse Phosphorylation-independent inhibition of parathyroid hormone receptor signaling by G protein-coupled receptor kinases PNAS, May 11, 1999; 96(10): 5476 - 5481. [Abstract] [Full Text] [PDF]

				G. Iguchi, Y. Okimura, T. Takahashi, I. Mizuno, M. Fumoto, Y. Takahashi, H. Kaji, H. Abe, and K. Chihara Cloning and Characterization of the 5'-Flanking Region of the Human Growth Hormone-releasing Hormone Receptor Gene J. Biol. Chem., April 23, 1999; 274(17): 12108 - 12114. [Abstract] [Full Text] [PDF]

				T. Massfelder and J.-J. Helwig Editorial: Parathyroid Hormone-Related Protein in Cardiovascular Development and Blood Pressure Regulation Endocrinology, April 1, 1999; 140(4): 1507 - 1510. [Full Text]

				Z. Huang, T. Bambino, Y. Chen, J. Lameh, and R. A. Nissenson Role of Signal Transduction in Internalization of the G Protein-Coupled Receptor for Parathyroid Hormone (PTH) and PTH-Related Protein Endocrinology, March 1, 1999; 140(3): 1294 - 1300. [Abstract] [Full Text]

				N. Malecz, T. Bambino, M. Bencsik, and R. A. Nissenson Identification of Phosphorylation Sites in the G Protein-Coupled Receptor for Parathyroid Hormone. Receptor Phosphorylation Is Not Required for Agonist-Induced Internalization Mol. Endocrinol., December 1, 1998; 12(12): 1846 - 1856. [Abstract] [Full Text]

				M. G. Barrett, G. S. Belinsky, and A. H. Tashjian Jr. A New Action of Parathyroid Hormone. RECEPTOR-MEDIATED STIMULATION OF EXTRACELLULAR ACIDIFICATION IN HUMAN OSTEOBLAST-LIKE SaOS-2 CELLS J. Biol. Chem., October 17, 1997; 272(42): 26346 - 26353. [Abstract] [Full Text] [PDF]

				G. Innamorati, H. Sadeghi, A. N. Eberle, and M. Birnbaumer Phosphorylation of the V2 Vasopressin Receptor J. Biol. Chem., January 24, 1997; 272(4): 2486 - 2492. [Abstract] [Full Text] [PDF]

				X. Zhen, L. Wei, Q. Wu, Y. Zhang, and Q. Chen Mitogen-activated Protein Kinase p38 Mediates Regulation of Chondrocyte Differentiation by Parathyroid Hormone J. Biol. Chem., February 9, 2001; 276(7): 4879 - 4885. [Abstract] [Full Text] [PDF]